ABSTRACT Core D (Pathology and Bio-Imaging Core), directed by Dr. Morrison, MD, will support this clinically oriented Program by providing comprehensive state-of-the-art evaluation of the immune status of tumor microenvironments (TME), using cutting-edge platforms for comprehensive immune profiling, cell and tissue analysis, imaging and image-analysis. Core D includes the following Aims: Aim 1: Provide infrastructure to collect, process, annotate and archive tumor samples and evaluate the immune status of the TME using comprehensive immune profiling. Our Tissue Procurement Service will help collect fresh, frozen and FFPE tumor specimens from the clinical trials of this Program and tissue banking protocols. OmniSeq and multiplex IHC subcore, will perform immunologic, genomic, and transcriptomic assessment of the immune status of the TME, using CLIA-certified Immune Report Card? (IRC), to determine: 1) transcript levels of 384 immune genes, 2) mutational burden of each tumor, 3) microsatellite instability, and 4) copy number gain of PD-L1 and PD-L2. We will also evaluate 5) on-treatment changes in TCR beta repertoire, to evaluate evidence of tumor-specific T cell response in the TME and 6) Patterns of expression of PD-1/PD-L1/PD-L2 system and COX2 system in relation to T cell infiltrate and clinical outcomes. Aim 2: Provide state-of-the-art immuno-fluorescence imaging and flow cytometry services to interrogate the spatial effects of chemokine modulating agents on immune cell influx, effector- and immuno-suppressive mechanisms in the TME. It will provide state-of-the-art multicolor flow and imaging cytometry, confocal laser scanning and multicolor fluorescence microscopy, combined with computer morphometry and image analysis. Aim 3 (Sub-Core D3): Perform small animal live imaging to determine the kinetics of tumor growth (bioluminescence) in differentially-treated mice and the kinetics of CTL traffic and accumulation in TME. Programmatic Role and Interactions. With Projects 1, 2 and 3 we will evaluate immune profiles of human colorectal, ovarian and melanoma tumors in the course of patient treatment with chemokine-modulating regimens and/or DC vaccines; determine changes in density, clonality and distribution of CTLs and other immune cells in human and mouse tumors treated by CKM in the absence and presence of vaccination and PD-1 blockade, and determine growth kinetics of the differentially treated and non-treated tumors in murine models. We will also determine the impact of treatments on vascular normalization; and formation of tumor- associated lymphoid structures implicated in epitope spreading and long term-treatment effects. Core D will interact with Core A for prioritization of services, with Core B to assure that proper design of studies and sound statistical evaluation of the results. We will work closely with Core C to ensure failsafe procurement of all specimens from the clinical trials, samples annotation and evaluation in context of clinical outcomes.